Abstract 16439: Proteomic Analysis of Atherogenic Chylomicron Remnants Isolated Selectively From Human Serum
Introduction: Many studies revealed the relationship between the enteric environment and development of cardiovascular diseases. Chylomicron remnants (CM-Rs), the hydrolyzed products from intestine-derived chylomicrons during the postprandial state, are known to be atherogenic, however it has been difficult to selectively isolate CM-Rs from other lipoproteins. We have, for the first time, established a method to selectively isolate CM-Rs from human serum using a monoclonal anti-apolipoprotein (apo)B-48 antibody, a major protein component of CM-Rs.
Hypothesis: We evaluated the proteomic profile of CM-Rs to elucidate their atherogenicity.
Methods: Postprandial serum samples of 8 healthy volunteers were collected 3 hours after a high-fat diet. Fractions containing CM-Rs and VLDL-sized particles (d<1.019 g/mL) were collected by ultracentrifugation and immunoaffinity isolation using Sepharose beads containing anti-apoB-48 or anti-apoB-100 mAbs. Serum lipoprotein profiles were analyzed by an HPLC method and the protein profile of isolated lipoproteins by shotgun proteomic analysis. Similar assessments were performed and protein mass was analyzed using postprandial intestinal lymph and intestinal mucosa in 8-week-old C57BL/6J mice.
Results: Isolated CMs from postprandial human serum contained lipoproteins with the size of large VLDL to small LDL, which strongly indicated the selectively separation of CM-Rs. These CM-Rs contained apoB, A2, apoD and apoH, complements (C3, C4 etc) and paraoxonase 1 (PON-1). Serum TG concentrations positively correlated with apoB-48, apoA5, apoC5 and PON-1. Spectrum counts of apoA4, apoA5, apoC1, apoM, clusterin, PON-1, prothrombin and fibrinogen were larger in CM-Rs than VLDL-remnants (VLDL-Rs) isolated anti-apoB-100 antibody. These proteins were also observed in mouse intestinal lymph with the increased concentrations of C3 and clusterin. These results strongly suggested that intestine-derived CM-Rs appearing during the postprandial period contained proteins related to inflammation and immune system.
Conclusions: CM-Rs contains more proteins involved in inflammation and atherosclerosis than VLDL-Rs, which may directly influence the atherogenesis during the postprandial period.
Author Disclosures: D. Masuda: Other Research Support; Modest; Fuji Rebio Company (as a joint research). T. Kobayashi: None. M. Sairyou: None. T. Okada: None. M. Koseki: None. T. Ohama: None. M. Nishida: None. Y. Sakata: None. S. Yamashita: Other Research Support; Modest; Fujirebio Inc. (as a joint research).
- © 2016 by American Heart Association, Inc.